GB/T 15805.5-2018Protocol of diagnosis for spring viraemia of carp (English PDF)

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard replaces GB/T 15805.5-2008 "Fish quarantine method Part 5. Hunchun viremia virus (SVCV)". versus Compared with GB/T 15805.5-2008, except for editorial changes, the main technical changes are as follows:

--- Added abbreviations;

--- Added enzyme-linked immunosorbent assay (ELISA) to detect hunchun viremia Method of antigen (springviraemiaofcarpvirus, SVCV);

---Incremental indirect immunofluorescent antibody test (IFAT) to detect SVCV Method of antigen;

--- Added real-time quantitative PCR (real-time PCR) method for detecting SVCV nucleic acid;

--- Increased the comprehensive judgment of springviraemiaofcarp (SVC). Please note that some of the contents of this document may involve patents. The issuing organization of this document is not responsible for identifying these patents. This standard was proposed by the Ministry of Agriculture and Rural Affairs of the People's Republic This standard is under the jurisdiction of the National Aquaculture Standardization Technical Committee (SAC/TC156). This standard was drafted. Ningbo Ocean and Fisheries Research Institute, National Aquatic Technology Extension Station, Beijing Customs, People's Republic of China, China Shekou Customs of the People's Republic of China. The main drafters of this standard. Wang Jianping, Zhang Lifeng, Jiang Yulin, Xu Shengwei, Gao Longying, Wu Xiongfei, Chen Aiping, Shen Weiliang. The previous versions of the standards replaced by this standard are.

---GB/T 15805.1-1995;

---GB/T 15805.5-2008. Hunchun viremia diagnosis procedure

1 Scope

GB/T 15805.5-2018 is the Chinese national standard on protocol of diagnosis for spring viraemia of carp, in the field of agriculture. The /T suffix marks it as a recommended standard: it is not compulsory by itself, but it becomes binding as soon as a contract, a tender or a customer specification calls it up - which in practice is how most foreign buyers meet it. It was issued on 28 December 2018 by the State Administration for Market Regulation; Standardization Administration of China, and has been in force since 1 July 2019. Classification: ICS 65.020.30, CCS B41. This page is published from the official record of the standard held by the Chinese standards administration: the identification, the dates, the classification and the issuing body are taken from there. The clause text, the tables and the numeric limits are in the document itself, which is delivered complete in English translation.

This standard specifies the sampling of viremia viremia, virus isolation and culture, RT-PCR, real-time quantitative RT-PCR, enzyme-linked immunosorbent assay Test, indirect immunofluorescent antibody test, comprehensive determination method. This standard applies to the epidemiological investigation, diagnosis, quarantine and epidemic monitoring of Hunchun viremia.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this article. Pieces. For undated references, the latest edition (including all amendments) applies to this document.

GB/T 6682 Analytical laboratory water specifications and test methods SC/T 7103 Technical specification for quarantine sampling of aquatic animals

3 Abbreviations

The following abbreviations apply to this document. CO. grass carp ovary cell line (grasscarpovarycelline) CPE. cytopathic effect EB. ethidium bromide (ethidiumbromide) ELISA. enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay) EPC. epithelioma cell line (epitheliomapapulosumcyprinicelline) FCS. fetal calf serum (fetalcalfserum) FHM. fat head muscle cell line (fatheadmonnowcelline) FITC. fluorescein isothiocyanate G protein. glycoprotein IFAT. indirect immunofluorescent antibody test (indirectimmunofluorescentantibodytest) RT-PCR. reverse transcription-polymerase chain reaction (reverse transcription polymerase chain reaction) SVC. spring viremia (springviraemiaofcarp) SVCV. springviraemiaofcarpvirus

4 Reagents and materials

4.1 Water. Meet the specifications of the first grade water in GB/T 6682; nucleic acid extraction and RT-PCR should use RNase-free deionized water.

4.2 SVCV reference strain. provided by the designated unit of the Ministry of Agriculture and Rural Areas of the People's Republic of China.

4.3 Fish cell lines. EPC, FHM, CO.

4.4 Fish cell growth solution. Prepared with Earle's balanced salt in M199 medium plus 10% fetal bovine serum (FCS).

4.5 Fish cell maintenance solution. Prepared with Earle's balanced salt in M199 medium plus 2 S.

4.6 Cell Digestion. See Appendix A, A.1.